Pathogen Identification and Biocontrol Screening of Root Rot of Cymbidium sinense

doi:10.11924/j.issn.1000-6850.casb2021-0239

Abstract

Abstract:

The paper aims to identify the pathogen of root rot and put forward green biological control measures. The pathogen of root rot of Cymbidium sinense was isolated by tissue isolation method, and identified by morphology and molecular biology. The pathogenicity was verified by Koch’s law. At the same time, 20 strains of bacteria were isolated from the roots and soil of wild healthy C. sinense by dilution coating method. Through plate confrontation culture, one strain of bacteria MJ5-1 was screened, which could antagonize two pathogens. The 16S rRNA gene sequence analysis showed that the antagonistic strain was Bacillus pumilus. The results showed that Fusarium solani was the pathogen of root rot of cultivated C. sinense in Mojiang. B. pumilus MJ5-1 could inhibit the growth of mycelium and spore of F. solani. This study could provide a theoretical basis for the diagnosis and pollution-free control of root rot of cultivated C. sinense.

Key words: root rot of Cymbidium sinense, tissue isolation method, pathogen, identification, antagonistic bacteria, screening

CLC Number:

S432.1

Tang Jie, Chen Jianxin, Nima Cimu, Wei Yuqian, Luo Zhui, Han Yuting, Lv Zejia, Ma Huancheng, Wu Jianrong. Pathogen Identification and Biocontrol Screening of Root Rot of Cymbidium sinense[J]. Chinese Agricultural Science Bulletin, 2021, 37(19): 128-133.

Figures/Tables 8

References 31

[1]	吴亚光. 兰花致命病一腐烂病[J]. 花木盆景(花卉园艺), 2004(3):26.
[2]	曹涤环. 兰花腐烂病的识别与防治[J]. 科普天地, 2015(8):8-8.
[3]	方中达. 植病研究方法(3版) [M]. 北京: 中国农业出版社, 1998: 124.
[4]	吴烨, 刘仲健. 兰花圆斑病的研究[J]. 华南农业大学学报, 1997(3):38-41.
[5]	Machado A R, Pinho D B, Pereira O L. Phylogeny, identification and pathogenicity of the Botryosphaeriaceae associated with collar and root rot of the biofuel plant Jatropha curcas in Brazil, with a description of new species of Lasiodiplodia[J]. Fungal Diversity, 2014, 67(1):231-247. doi: 10.1007/s13225-013-0274-1 URL
[6]	刘仲健, 罗焕亮, 陈伟元, 等. 兰花叶枯病的病原鉴定[J]. 华南农业大学学报, 1997(1):31-33.
[7]	邵力平, 沈瑞祥, 张素轩. 真菌分类学[M]. 北京: 中国农业出版社, 1984:79-80.
[8]	李欣, 崔秀明, 刘迪秋, 等. 三七根腐病菌尖孢镰刀菌的Real-timePCR检测方法[J]. 华北农学报, 2019(增刊). 45(4):224-330.
[9]	郑琪, 徐秉良, 薛应钰, 等. 杏采后病害病原拮抗菌的分离筛选及鉴定[J]. 植物保护, 2013, 39(4):34-39,71.
[10]	Foldes T, Banhegyi I, Herpai Z, et al. Isolation of Bacillus strains from the rhizosphere of cereals and in vitro screening for antagonism against phytopathogenic, food-borne pathogenic and spoilage micro-oganisms[J]. Journal of Applied Microbiology, 2000, 89(5):840-846. doi: 10.1046/j.1365-2672.2000.01184.x URL
[11]	Voisard C, Keel C, Haas D, et al. Cyanide production by Pseudomonas fluorescens helps suppress black root rot of tobacco under gnotobiotic conditions[J]. The EMBO Journal, 1989, 8(2):351-358. doi: 10.1002/embj.1989.8.issue-2 URL
[12]	Ahmed A S, Pérez ánchez C, Egea C, et al. Evaluation of Trichoderma harzianum for controlling root rot caused by Phytophthora capsici in pepper plants[J]. Plant Pathology, 2006, 48(1):58-65. doi: 10.1046/j.1365-3059.1999.00317.x URL
[13]	牛慧芹, 刘春辉, 沈检龙, 等. 玉米大斑病生防细菌的筛选、鉴定及其抑制作用[J]. 中国农学通报, 2014, 30(28):275-279.
[14]	连彩, 郭晓军, 朱宝成, 等. 兰花枯萎病拮抗细菌的筛选与鉴定[J]. 华北农学报, 2012(2):226-229.
[15]	Wang R, Dong L L, Xu J, et al. Identification and prevention of root rot pathogen in model of ginseng cultivated in farmlands[J]. China Journal of Chinese Materia Medica, 2016, 41(10):1787-1791. doi: 10.4268/cjcmm20161004 pmid: 28895321
[16]	Chang X, Dai H, Wang D, et al. Identification of Fusarium species associated with soybean root rot in Sichuan Province, China[J]. European Journal of Plant Pathology, 2018, 151(5):1-15.
[17]	Siddiqui I A, Ehetshamul-Haque S, Shaukat S S. Use of Rhizobacteria in the Control of Root Rot-Root Knot Disease Complex of Mungbean[J]. Journal of Phytopathology, 2001, 149.
[18]	Dawar S, Hayat S, Anis M, et al. Effect of seed coating material in the efficacy of microbial antagonists for the control of root rot fungi on Okra and Sunflower[J]. Pakistan Journal of Botany, 2014, 40(3):1269-1278.
[19]	Shen T, Wang C, Yang H, et al. Identification, solid-state fermentation and biocontrol effects of Streptomyces hygroscopicus B04 on strawberry root rot[J]. Applied Soil Ecology, 2016, 103:36-43. doi: 10.1016/j.apsoil.2016.02.016 URL
[20]	Sun W M, Ma Y N, Yin Y J, et al. Effects of Essential Oils from Zingiberaceae Plants on Root-Rot Disease of Panax notoginseng[J]. Molecules, 2018, 23(5):1021. doi: 10.3390/molecules23051021 URL
[21]	Wang R Y, Gao B, Li X H, et al. First Report of Fusarium solani Causing Fusarium Root Rot and Stem Canker on Storage Roots of Sweet Potato in China[J]. Plant Disease, 2014, 98(1):160. doi: 10.1094/PDIS-06-13-0651-PDN pmid: 30708580
[22]	Chatterton S, Bowness R, Harding M W. First Report of Root Rot of Field Pea Caused by Aphanomyces euteiches in Alberta, Canada[J]. Plant Disease, 2015, 99(2):288. doi: 10.1094/PDIS-09-14-0905-PDN pmid: 30699595
[23]	Wang Q, Buxa S V, Furch A, et al. Insights Into Triticum aestivum Seedling Root Rot Caused by Fusarium graminearum[J]. Molecular Plant-Microbe Interactions, 2015, 28(12).
[24]	雷恒树, 华战迎, 范灵姣, 等. 北京地区草莓枯萎病病原的鉴定与防治[J]. 中国农业大学学报, 2019, 24(6):66-72.
[25]	刘香萍, 敬学敏, 闫红秀, 等. 尖孢镰刀菌的分离鉴定与不同木薯品种芽期抗病性[J]. 植物保护, 2019, 45(4):229-235.
[26]	赵彩呈, 洪英娣, 刘丽, 等. 丽江点重楼和白芨根腐病病原鉴定[J]. 贵州农业科学, 2019, 47(5):28-30.
[27]	周盈, 陈琳, 柴鑫莉, 等. 魔芋内生拮抗细菌的分离及其抗菌物质特性研究[J]. 微生物学报, 2007, 47(6):1076-1079.
[28]	陈琼珍, 吴兴泉. 枯草芽孢杆菌对有害真菌的生防作用及最佳发酵条件研究[J]. 河南工业大学学报:自然科学版, 2011, 32(5):66-70.
[29]	高芬, 任小霞, 王梦亮, 等. 中草药根腐病及其微生物防治研究进展[J]. 中国中药杂志, 2015, 40(21):4122-4126.
[30]	王英国, 王军华, 权春善, 等. 解淀粉芽孢杆菌抗菌活性物质的分离纯化及抑菌活性研究[J]. 中国生物工程杂志, 2007(12):41-45.
[31]	勾长龙, 王雨琼, 孙朋, 等. 人参根腐病拮抗菌的筛选、鉴定及其抑菌活性[J]. 食品科学, 2015, 36(19):143-147.

供试菌株F. Solani	抑菌直径/mm	无菌滤液抑菌率/%
MJ1-1	30.6±1.2b	96.68±0.214bc
MJ5-1	43.6±1.2a	100.68±1.13ab
MJ7-2	29.6±1.2b	97.68±1.2bc

供试菌株F. Solani	抑菌直径/mm	无菌滤液抑菌率/%
MJ1-1	30.6±1.2b	96.68±0.214bc
MJ5-1	43.6±1.2a	100.68±1.13ab
MJ7-2	29.6±1.2b	97.68±1.2bc